Department of Animal Developmental Biology, Faculty of Biological Sciences, University of Wroclaw, Sienkiewicza 21, 50-335 Wroclaw, Poland.
Hirszfeld Institute of Immunology and Experimental Therapy, the Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wroclaw, Poland.
Cells. 2020 Jun 26;9(6):1562. doi: 10.3390/cells9061562.
Hspb8 is a member of the small heat shock protein (sHSP) family. Its expression is known to be upregulated under heat shock. This protein interacts with different partners and can, therefore, be involved in various processes relevant to tissue integrity and functioning. In humans, mutations in the gene encoding Hspb8 can lead to the development of various diseases such as myopathies and neuropathies. In our study, we aimed to perform an in-depth characterization of zebrafish Hspb8 during zebrafish development. We applied techniques such as RT-qPCR, Western blot, immunofluorescence, co-immunoprecipitation, LC-MS, and morpholino-mediated knockdown. We broadened the knowledge regarding zebrafish expression during development under normal and heat shock conditions as well as its tissue- and subcellular-specific localization. A co-IP analysis allowed us to conclude that zebrafish Hspb8 can interact with proteins such as Bag3 and Hsc70, which are crucial for formation of an autophagy-inducing complex. We also demonstrated that morpholino-mediated knockdown has an impact on zebrafish embryos' morphology, muscle ultrastructure, and motility behavior. Our research provides a valuable resource for the potential use of the zebrafish as a model for studying pathological conditions associated with disorders.
Hspb8 是小分子热休克蛋白 (sHSP) 家族的成员。已知其表达在热休克下上调。这种蛋白质与不同的伴侣相互作用，因此可以参与与组织完整性和功能相关的各种过程。在人类中，编码 Hspb8 的基因突变可导致各种疾病的发展，如肌病和神经病。在我们的研究中，我们旨在对斑马鱼发育过程中的 Hspb8 进行深入表征。我们应用了 RT-qPCR、Western blot、免疫荧光、共免疫沉淀、LC-MS 和基于 morpholino 的敲低等技术。我们拓宽了关于正常和热休克条件下斑马鱼发育过程中表达以及其组织和亚细胞特异性定位的知识。共免疫沉淀分析使我们能够得出结论，即斑马鱼 Hspb8 可以与 Bag3 和 Hsc70 等蛋白质相互作用，这些蛋白质对于形成诱导自噬的复合物至关重要。我们还证明，基于 morpholino 的敲低会对斑马鱼胚胎的形态、肌肉超微结构和运动行为产生影响。我们的研究为潜在使用斑马鱼作为研究与疾病相关的病理条件的模型提供了有价值的资源。
